Methods of obtaining compositions enriched for hematopoietic stem cells, compositions derived therefrom and methods of use thereof

Abstract

A method for obtaining human hematopoietic stem cells is provided by enrichment for stem cells using a novel stem cell marker. Compositions enriched for stem cells are obtained using novel antibody HCC-1 which is specific for a CD59 epitope highly accessible on stem cells, and less accessible or absent on mature cells. The enriched population of cells derived from these methods are also provided.

What is claimed is: 
     
       1. A method of obtaining a composition substantiaily enriched in subpopulation of hematopoiefic stem cells comprising: (a) obtaining, a source of cells comprising human hematopoietic cells;   (b) isolating a population of human hematopoietic cells from the source;   (c) contacting the population of human hematopoietic cells with a monoclonal antibody specific for the marker HCC-1; and   (d) separating the subpopulation that is specifically bound by the monoclonal antibody, thereby obtaining a composition substantially enriched in a subpopulation of human hematopoietic stem cells.   
     
     
       2. The method accoraing to claim 1 wherein the step of isolating a population of hematopoietic cells is by selecting the cells for expression of at least one additional marker associated with stem cells or by physical separation means. 
     
     
       3. The method according to claim 2 wherein the additional marker is selected from the group consisting of CD34, Thy-1 + , rho l , and C-Kit + . 
     
     
       4. A composition comprising a substantially enriched subpopulation of cells expressing HCC-1 obtained by the method according to claim 3. 
     
     
       5. A composition comprising a substantially enriched subpopulation of cells expressing HCC-1 obtained by the method according to claim 2. 
     
     
       6. A composition comprising a substantially enriched subpopulation of cells expressing HCC-1 obtained by the method according to claim 1. 
     
     
       7. The method according to claim 1 wherein the step of isolating a population of hematopoietic cells is by selecting cells for the absence of at least one lineage specific (LIN - ) marker. 
     
     
       8. The method according to claim 7 wherein the lineage specific marker is selected from the group consisting of CD14 and CD15, CD38, HLA-DR, CD71, and CD33. 
     
     
       9. A composition comprising a substantially enriched Subpopulation of cells expressing HCC-1 obtained by the method according to claim 8. 
     
     
       10. The method according to claim 7 wherein the lineage specific marker is selected from the group consisting of CD2, CD16, CD19, and glycophorin A. 
     
     
       11. A composition comprising a substantially enriched subpopulation of cell's expressing HCC-1 obtained by the method according to claim 5. 
     
     
       12. A composition comprising a substantially enriched subpopulation of cells expressing HCC-1 obtained by the method according to claim 7. 
     
     
       13. The method according to claim 7 wherein the step of isolating a population of human hematopoietic cells further comprises selecting the cells for the expression of at least one additional marker associated with stem cells. 
     
     
       14. A composition comprising a substantially enriched subpopulation of cells expressing HCC-1 obtained by the method according to claim 13. 
     
     
       15. The method according to claim 13 wherein the additional marker is selected from the group consisting of CD34, Thy-1 + , rho lo  and C-Kit + . 
     
     
       16. A composition comprising a substantially enriched subpopulation of cells expressing HCC-1 obtained by the method according to claim 15. 
     
     
       17. The method according to claim 13 whereto the lineage specific marker is selected from the group consisting of CD14, CD15, CD38, HLA-DR, CD71 and CD33. 
     
     
       18. A composition comprising a substantially enriched subpopulation of cells expressing HCC-1 obtained by the method according to claim 17. 
     
     
       19. The method according to claim 13 wherein the lineage specific marker is selected from the group consisting of CD2, CD16, CD19 and glycophorin A. 
     
     
       20. A composition comprising a substantially enriched subpopulation of cells expressing HCC-1 obtained by the method according to claim 19. 
     
     
       21. The method according to claim 1, where, step (c) is performed prior to step (b). 
     
     
       22. The composition substantially enriched in a subpopulation of hematopoietic stem cells obtained by the method of claim 21. 
     
     
       23. The hybridoma cell line designated ATCC Accession No. HB 11729. 
     
     
       24. The monoclonal antibody produced by the hybridoma cell line of claim 23. 
     
     
       25. An antibody that specifically binds to the epitiope that is specifically bound by the monoclonal antibody of claim 24. 
     
     
       26. A fragment of the antibody of claim 24 that includes the variable region of at least one heavy or light chain.